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Nucleic Acids Symposium Series 2007 51(1):3-4; doi:10.1093/nass/nrm002
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© 2007 Oxford University Press

Chemical synthesis of a very long RNA oligomer, a 110mer precursor-miRNA candidate, with 2-cyanoethoxymethyl (CEM) as the 2'-O-protecting group

Hirofumi Masuda*, Yoshinobu Shiba, Naoki Watanabe, Kazuchika Takagaki, Kouichi Ishiyama, Tadaaki Ohgi and Junichi Yano

Discovery Research Laboratories, Nippon Shinyaku Co. Ltd., 3-14-1 Sakura, Tsukuba City, Ibaraki 305-0003, Japan

* h.toyoshima{at}po.nippon-shinyaku.co.jp

Abstract

A long RNA oligomer, a 110mer with the sequence of a precursor-miRNA candidate, has been chemically synthesized in a single synthesizer run by means of standard automated phosphoramidite chemistry. The synthetic method involved the use of 2-cyanoethoxymethyl (CEM), a 2'-hydroxyl protecting group recently developed in our laboratory. We confirmed the identity of the synthetic 110mer by MALDI-TOF mass spectrometry, as well as HPLC, electrophoretic methods, RNase-digestion experiments, and its in vitro gene-silencing activity. The chemical synthesis of RNA oligomers of more than 100 nucleotides, which has until now been extremely difficult, can be practically realized by the CEM method.


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